Effects of heavy ions on inactivation and DNA double strand breaks in Deinococcus radiodurans R1.

Inactivation and double strand break (dsb) induction after heavy ion irradiation were studied in stationary phase cells of the highly radiation resistant bacterium Deinococcus radiodurans R1. There is evidence that the radiation sensitivity of this bacterium is nearly independent on energy in the range of up to 15 MeV/u for lighter ions (Ar). The responses to dsb induction for charged particles show direct relationship between increasing radiation dose and residual intact DNA.     

Double strand breaks in the DNA of Bacillus subtilis cells irradiated by heavy ions.

Cells of Bacillus subtilis strain TKJ 8431 in stationary phase were irradiated with X-rays (150 kV at DLR) or heavy ions (Ne, Ar, Pb with residual energies between 3 and 15 MeV/u at GSI). The action cross section for the formation of double strand breaks in the DNA of the irradiated cells follows a similar dependence on mass and energy of the ions as has been found for various biological endpoints, e.g. inactivation, mutagenesis and repair efficacy.     

Inactivation, mutation induction and repair in Bacillus subtilis spores irradiated with heavy ions.

Studies on the response of bacterial spores to accelerated heavy ions (HZE particles) help in understanding problems of space radiobiology and exobiology. Layers of spores of Bacillus subtilis strains, differing in repair capabilities, were irradiated with accelerated boron, carbon and neon ions of linear energy transfer (LET) values up to 14000 MeV cm2/g. Inactivation as measured by loss of colony forming ability and induction of mutations as measured by reversion to histidine prototrophy and resistance to 150 micrograms/ml sodium azide were tested, as well as the influence of repair processes on these effects. For inactivation, the cross-sectional values sigma plotted as a function of LET follow a saturation curve. The plateau, which is reached around a LET of 2000 MeV cm2/g, occurs at 2.5 x 10(-9) cm2, a value in good agreement with the dimensions of the spore protoplast. Lethal damage produced at LET values < 2000 MeV cm2/g is reparable. Recombination repair is more effective than excision repair. At higher LET values, lethal damage could not be reconstituted by the repair mechanisms studied. In addition, at these high LET values, the frequency of induced mutations was drastically decreased. The data support the assumption of at least two qualitatively different types of lesion, depending on the LET of the affecting heavy ion.     

Cell inactivation, repair and mutation induction in bacteria after heavy ion exposure: results from experiments at accelerators and in space.

To understand the mechanisms of accelerated heavy ions on biological matter, the responses of spores of B. subtilis to this structured high LET radiation was investigated applying two different approaches. 1) By the use of the Biostack concept, the inactivation probability as a function of radial distance to single particles' trajectory (i.e. impact parameter) was determined in space experiments as well as at accelerators using low fluences of heavy ions. It was found that spores can survive even a central hit and that the effective range of inactivation extends far beyond impact parameters where inactivation by delta-ray dose would be effective. Concerning the space experiment, the inactivation cross section exceeds those from comparable accelerator experiments by roughly a factor of 20. 2) From fluence effect curves, cross sections for inactivation and mutation induction, and the efficiency of repair processes were determined. They are influenced by the ions characteristics in a complex manner. According to dependence on LET, at least 3 LET ranges can be differentiated: A low LET range (app. < 200 keV/micrometers), where cross sections for inactivation and mutation induction follow a common curve for different ions and where repair processes are effective; an intermediate LET range of the so-called saturation cross section with negligible mutagenic and repair efficiency; and a high LET range (>1000 keV/micrometers) where the biological endpoints are majorly dependent on atomic mass and energy of the ion under consideration.     

Quantitative interpretation of heavy ions effects: models for the biological effects of heavy ions.

Heavy ions are an important part of space radiation. Although they contribute only about 1 percent in number the fraction in terms of energy deposited is much higher. Also the quality of radiation is different from the other components since the LET is generally quite high. This poses the problem of Relative Biological Effectiveness (RBE). It is considerably more important in space than on earth because shielding measures are costly and sometimes not even feasible. Radiation hazards appear to be the limiting factor In long term space flights and their evaluation constitutes a major task. There is still no general agreement about RBE of earthbound radiation, and even less concerning the biological weighting of very heavy and very energetic ions in space. Because of the lack of experimental data--particularly for risk estimates in humans-- theoretical approaches may be very helpful in this respect and provide the only means to judge the radiation protection situation in outer space. In order to be useful careful checks of their consistency are necessary. This paper summarizes some of the more common approaches in a critical manner. The unhappy conclusion at the end will be that at present it is not possible to understand even heavy ion action on survival quantitatively with an acceptable precision.     

Biophysical effect of cosmic heavy ions of distinct LET-classes in a plant model system.

Results presented from recent space flight BION 9 show biological effects of different LET-classes of HZE-particles in different target regions of the seed (meristem and the whole embryo) of Arabidopsis thaliana (L.) Heyhn. HZE-one hit events and non-hit events, i.e. only hit by the low-LET background radiation, and their combined effects on the biological damage endpoint lethality are distinguished. This procedure is opening the opportunity of an approach to comparative studies of the biological effects induced by cosmic HZE-particles of different LET-ranges interacting in the complex cosmic radiation spectrum and with other space flight conditions.     

LET dependence of cell death, mutation induction and chromatin damage in human cells irradiated with accelerated carbon ions.

We investigated the LET dependence of cell death, mutation induction and chromatin break induction in human embryo (HE) cells irradiated by accelerated carbon-ion beams. The results showed that cell death, mutation induction and induction of non-rejoining chromatin breaks detected by the premature chromosome condensation (PCC) technique had the same LET dependence. Carbon ions of 110 to 124keV/micrometer were the most effective at all endpoints. However, the number of initially induced chromatin breaks was independent of LET. About 10 to 15 chromatin breaks per Gy per cell were induced in the LET range of 22 to 230 keV/micrometer. The deletion pattern of exons in the HPRT locus, analyzed by the polymerase chain reaction (PCR), was LET-specific. Almost all of the mutants induced by 124 keV/micrometer beams showed deletion of the entire gene, while all mutants induced by 230keV/micrometer carbon-ion beams showed no deletion. These results suggest that the difference in the density distribution of carbon-ion track and secondary electron with various LET is responsible for the LET dependency of biological effects.     

Effect of track structure and radioprotectors on the induction of oncogenic transformation in murine fibroblasts by heavy ions

The oncogenic potential of high-energy ⁵⁶Fe particles (1 GeV/nucleon) accelerated with the Alternating Gradient Synchrotron at the Brookhaven National Laboratory was examined utilizing the mouse C3H 10T1/2 cell model. The dose-averaged LET for high-energy ⁵⁶Fe is estimated to be 143 keV/μm with the exposure conditions used in this study. For ⁵⁶Fe ions, the maximum relative biological effectiveness (RBE_max) values for cell survival and oncogenic transformation were 7.71 and 16.5 respectively. Compared to 150 keV/μm ⁴He nuclei, high-energy ⁵⁶Fe nuclei were significantly less effective in cell killing and oncogenic induction. The prostaglandin E₁ analog misoprostol, an effective oncoprotector of C3H 10T1/2 cells exposed to X rays, was evaluated for its potential as a radioprotector of oncogenic transformation with high-energy ⁵⁶Fe. Exposure of cells to misoprostol did not alter ⁵⁶Fe cytotoxicity or the rate of ⁵⁶Fe-induced oncogenic transformation.     

Effects of heavy ions on visual function and electrophysiology of rodents: the ALTEA-MICE project.

ALTEA-MICE will supplement the ALTEA project on astronauts and provide information on the functional visual impairment possibly induced by heavy ions during prolonged operations in microgravity. Goals of ALTEA-MICE are: (1) to investigate the effects of heavy ions on the visual system of normal and mutant mice with retinal defects; (2) to define reliable experimental conditions for space research; and (3) to develop animal models to study the physiological consequences of space travels on humans. Remotely controlled mouse setup, applied electrophysiological recording methods, remote particle monitoring, and experimental procedures were developed and tested. The project has proved feasible under laboratory-controlled conditions comparable in important aspects to those of astronauts' exposure to particle in space. Experiments are performed at the Brookhaven National Laboratories [BNL] (Upton, NY, USA) and the Gesellschaft für Schwerionenforschung mbH [GSI]/Biophysik (Darmstadt, FRG) to identify possible electrophysiological changes and/or activation of protective mechanisms in response to pulsed radiation. Offline data analyses are in progress and observations are still anecdotal. Electrophysiological changes after pulsed radiation are within the limits of spontaneous variability under anesthesia, with only indirect evidence of possible retinal/cortical responses. Immunostaining showed changes (e.g. increased expression of FGF2 protein in the outer nuclear layer) suggesting a retinal stress reaction to high-energy particles of potential relevance in space.     

Physical events in the track structure of heavy ions and their relation to alterations of biomolecules.

Heavy charged particles interacting with biological cells can produce a wide variety of different physical, chemical and biological consequences. A rigorous identification of relevant chemical and biological alterations of biomolecules in cells, however, is still lacking and, thus, it is difficult to identify the potential biological importance of different early physical events. In addition, due to experimental and theoretical problems also little is known about the details of energy transfer, -absorption and -decay from projectiles to atoms/molecules in condensed targets; this is particularly true for not completely stripped heavy ions. Nevertheless, one might conclude from available data that higher densities of physical energy absorption events have a significantly higher probability to lead to qualitatively more severe biochemical alterations as regards the induction of DNA double strand breaks and of chromatin damage. It is not very likely that energy migration along the DNA molecule in biological cells over long distances plays a significant role as contributor to these biological radiation effects.     

The effect of track structure on cell inactivation and chromosome damage at a constant LET of 120 keV/micrometer.

The influence of track structure on chromosome damage and cell inactivation are being investigated. Plateau-phase normal human fibroblast cultures were irradiated with gamma rays, and He, Ne and Ar ions. Particle velocities were chosen so that all beams had an LET of 120 keV/micrometer. In this constant-LET experimental design, the radial distribution of excitations and ionizations about the particle track is the most significant variable. Using premature chromosome condensation, chromatin breaks were measured at two time points, promptly after irradiation and after a prolonged incubation to allow for repair. These measurements give an indication of both initial chromosomal damage and also residual damage that is either not repaired or is misrepaired. Survival was measured under the same conditions. Results indicate that the RBEs for both cell inactivation and, to a lesser extent, chromosome damage decrease as particle energy increases.     

Damage to the photoreceptor cells of the rabbit retina from 56Fe ions: effect of age at exposure, 1.

Optic and proximate tissues of New Zealand white (NZW) rabbits at ages (approximately 3.5 years) near the middle of their median lifespan (5-7 years) were given 0.5-3.5 Gy of 465 MeV u-1 56Fe ions in the Bragg plateau region of energy deposition at a linear energy transfer (LET infinity) of 220 +/- 31 keV micrometer-1. Dose-dependent losses of retinal photoreceptor cells (rods) occurred until 1-2 years after irradiation, the period of this interim report. Similar cumulative losses of photoreceptor cells were seen during the period 1-2 years post-irradiation for rabbits given comparable exposures when young (6-9 weeks old). Since losses of photoreceptor cells at early times had not been determined previously, the current experiment, which was designed to simulate the responses of mature astronauts, redressed that deficiency.     

Preliminary results of space experiment: Implications for the effects of space radiation and microgravity on survival and mutation induction in human cells

In view of the concern for the health of astronauts that may one day journey to Mars or the Moon, we investigated the effect that space radiation and microgravity might have on DNA damage and repair. We sent frozen human lymphoblastoid TK6 cells to the International Space Station where they were maintained under frozen conditions during a 134-day mission (14 November 2008 to 28 March 2009) except for an incubation period of 8 days under 1G or μG conditions in a CO₂ incubator. The incubation period started after 100 days during which the cells had been exposed to 54 mSv of space radiation. The incubated cells were then refrozen, returned to Earth, and compared to ground control samples for the determination of the influence of microgravity on cell survival and mutation induction. The results for both varied from experiment to experiment, yielding a large SD, but the μG sample results differed significantly from the 1G sample results for each of 2 experiments, with the mean ratio of μG to 1G being 0.55 for the concentration of viable cells and 0.59 for the fraction of thymidine kinase deficient (TK⁻) mutants. Among the mutants, non-loss of zygosity events (point mutations) were less frequent (31%) after μG incubation than after 1G incubation, which might be explained by the influence of μG on cellular metabolic or physiological function. Additional experiments are needed to clarify the effect of μG interferes on DNA repair.     

Significant proton events of solar cycle 22 and a comparison with events of previous solar cycles.

The sun has produced several high energy and large fluence solar proton events during solar cycle 22. This recent activity is similar to activity that occurred in the 19th solar cycle before the advent of routine space measurements. In a review of the recent events and a comparison with significant solar proton events of previous solar cycles, it appears that the 20th and 21st solar cycles were deficient in the total fluence of solar particles as detected at the earth. Therefore, when models of maximum solar proton fluxes to be encountered for deep space missions are developed, solar proton data acquired during the present solar cycle should be incorporated.     

Solar-interplanetary circumstances associated with the major events in March and June 1991 and comparison with similar events of previous solar cycles.

The solar activity and geomagnetic storm events of March and June 1991 were associated with the appearance of an enhanced particle flux in the trapped radiation belts as discovered by the CRRES satellite observations and later measured by shuttle radiation detectors. The solar-interplanetary conditions associated with these events appear to be a major sequence of activity near the sun's central meridian generating powerful fast interplanetary shocks resulting in major perturbations to the magnetosphere. The solar-interplanetary events in 1991 are discussed and compared to similar activity in the past such as the events in February 1986, August 1972, July 1961, November 1960, and July 1959.     

The conceptual design of a hybrid life support system based on the evaluation and comparison of terrestrial testbeds.

This report summarizes a trade study of different options of a bioregenerative Life Support System (LSS) and a subsequent conceptual design of a hybrid LSS. The evaluation was based mainly on the terrestrial testbed projects MELISSA (ESA) and BIOS (Russia). In addition, some methods suggested by the Advanced Life Support Project (NASA) were considered. Computer models, including mass flows were established for each of the systems with the goal of closing system loops to the extent possible. In order to cope with the differences in the supported crew size and provided nutrition, all systems were scaled for supporting a crew of six for a 780 day Mars mission (180 days transport to Mars; 600 days surface period) as given in the NASA Design Reference Mission Scenario [Hoffman, S.J., Kaplan, D.L. Human exploration of Mars: the Reference Mission of the NASA Mars Exploratory Study, 1997]. All models were scaled to provide the same daily allowances, as of calories, to the crew. Equivalent System Mass (ESM) analysis was used to compare the investigated system models against each other. Following the comparison of the terrestrial systems, the system specific subsystem options for Food Supply, Solid Waste Processing, Water Management and Atmosphere Revitalization were evaluated in a separate trade study. The best subsystem technologies from the trade study were integrated into an overall design solution based on mass flow relationships. The optimized LSS is mainly a bioregenerative system, complemented by a few physico-chemical elements, with a total ESM of 18,088 kg, which is about 4 times higher than that of a pure physico-chemical LSS, as designed in an earlier study.     

Effects of 40Ar and 56Fe ions on retinal photoreceptor cells of the rabbit: implications for manned missions to Mars.

Losses of photoreceptor cells (rods) from the retinas of New Zealand white (NZW) rabbits were detectable within 2 years after localized acute irradiation of optic and proximal tissues with > or = 7 Gy of 530 MeV u-1 40Ar ions or > or = 2 Gy of 465 MeV u-1 56Fe ions in the Bragg plateau region of energy deposition. Those limits were determined only from an analysis of variance of dose groups because the shapes of the dose response curves at early post-irradiation times are not known, a concern being addressed by experiments in progress. Losses of photoreceptor cells for the period 0.5-2.5 years post-irradiation, determined by provisional linear regression analysis, were approximately 1.7% Gy-1 and 2.5% Gy-1 for 40Ar and 56Fe ions, respectively.     

Bottom side profiles for two close stations at the southern crest of the EIA: Differences and comparison with IRI-2012 and NeQuick2 for low and high solar activity

Bottom side electron density profiles for two stations at the southern crest of the Equatorial Ionization Anomaly (EIA), São José dos Campos (23.1°S, 314.5°E, dip latitude 19.8°S; Brazil) and Tucumán (26.9°S, 294.6°E, dip latitude 14.0°S; Argentina), located at similar latitude and separated by only 20° in longitude, have been compared during equinoctial, winter and summer months under low (year 2008, minimum of the solar cycle 23/24) and high solar activity (years 2013–2014, maximum of the solar cycle 24) conditions. An analysis of parameters describing the bottom side part of the electron density profile, namely the peak electron density NmF2, the height hmF2 at which it is reached, the thickness parameter B₀ and the shape parameter B₁, is carried out. Further, a comparison of bottom side profiles and F-layer parameters with the corresponding outputs of IRI-2012 and NeQuick2 models is also reported. The variations of NmF2 at both stations reveal the absence of semi-annual anomaly for low solar activity (LSA), evidencing the anomalous activity of the last solar minimum, while those related to hmF2 show an uplift of the ionosphere for high solar activity (HSA). As expected, the EIA is particularly visible at both stations during equinox for HSA, when its strength is at maximum in the South American sector. Despite the similar latitude of the two stations upon the southern crest of the EIA, the anomaly effect is more pronounced at Tucumán than at São José dos Campos. The differences encountered between these very close stations suggest that in this sector relevant longitudinal-dependent variations could occur, with the longitudinal gradient of the Equatorial Electrojet that plays a key role to explain such differences together with the 5.8° separation in dip latitude between the two ionosondes. Furthermore at Tucumán, the daily peak value of NmF2 around 21:00 LT during equinox for HSA is in temporal coincidence with an impulsive enhancement of hmF2, showing a kind of “elastic rebound” under the action of the EIA. IRI-2012 and NeQuick2 bottom side profiles show significant deviations from ionosonde observations. In particular, both models provide a clear underestimation of the EIA strength at both stations, with more pronounced differences for Tucumán. Large discrepancies are obtained for the parameter hmF2 for HSA during daytime at São José dos Campos, where clear underestimations made by both models are observed. The shape parameter B₀ is quite well described by the IRI-2012 model, with very good agreement in particular during equinox for both stations for both LSA and HSA. On the contrary, the two models show poor agreements with ionosonde data concerning the shape parameter B₁.